Zampanolide Binding to Tubulin Indicates Cross-Talk of Taxane Site with Colchicine and Nucleotide Sites
| Autor: | Michel Jordi, John H. Miller, J. Fernando Díaz, Benet Pera, Grégory Menchon, Juan Estévez Gallego, José Manuel Andreu, Javier Rodríguez-Salarichs, Didier Zuwerra, Andrea E. Prota, Enrique Calvo, Gonzalo Sáez-Calvo, Jessica J. Field, Karl-Heinz Altmann |
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| Přispěvatelé: | Ministerio de Economía y Competitividad (España), Cancer Society of New Zealand, Wellington Medical Research Foundation |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Bridged-Ring Compounds Stereochemistry Dimer Pharmaceutical Science macromolecular substances Biology Ligands Microtubules Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound Tubulin Drug Discovery medicine Colchicine Animals Humans Nucleotide Binding site Pharmacology chemistry.chemical_classification Biological Products Binding Sites Nucleotides Organic Chemistry Ligand (biochemistry) 030104 developmental biology Complementary and alternative medicine chemistry Mechanism of action Covalent bond biology.protein Molecular Medicine Cattle Taxoids Macrolides medicine.symptom |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname |
| ISSN: | 1520-6025 |
| Popis: | 41 p.-8 fig.-2 tab. Field, Jessica J. et al. The marine natural product zampanolide and analogues thereof constitute a new chemotype of taxoid site microtubule-stabilizing agents with a covalent mechanism of action. Zampanolide-ligated tubulin has the switch-activation loop (M-loop) in the assembly prone form and, thus, represents an assembly activated state of the protein. In this study, we have characterized the biochemical properties of the covalently modified, activated tubulin dimer, and we have determined the effect of zampanolide on tubulin association and the binding of tubulin ligands at other binding sites. Tubulin activation by zampanolide does not affect its longitudinal oligomerization but does alter its lateral association properties. The covalent binding of zampanolide to β-tubulin affects both the colchicine site, causing a change of the quantum yield of the bound ligand, and the exchangeable nucleotide binding site, reducing the affinity for the nucleotide. While these global effects do not change the binding affinity of 2-methoxy-5-(2,3,4-trimethoxyphenyl)-2,4,6-cycloheptatrien-1-one (MTC) (a reversible binder of the colchicine site), the binding affinity of a fluorescent analogue of GTP (Mant-GTP) at the nucleotide E-site is reduced from 12 ± 2 × 105 M-1 in the case of unmodified tubulin to 1.4 ± 0.3 × 105 M-1 in the case of the zampanolide tubulin adduct, indicating signal transmission between the taxane site and the colchicine and nucleotide sites of β-tubulin. This work was supported in part by grants BFU2016-75319-R (AEI/FEDER, UE) from Ministerio de Economia y Competitividad and the Cancer Society of New Zealand, and the Wellington Medical Research Foundation (JJF & JHM). The authors acknowledge networking contribution by the COST Action CM1407 “Challenging organic syntheses inspired by nature - from natural products chemistry to drug discovery”. The CNIC is supported by the Ministerio de Ciencia e Innovación and the Fundación Pro CNIC. |
| Databáze: | OpenAIRE |
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